Figure 1From: Androgen up-regulates vascular endothelial growth factor expression in prostate cancer cells via an Sp1 binding siteAndrogen regulates VEGF expression in LNCaP and 22Rv1 prostate cancer cells. (A) 22Rv1 cells were serum starved overnight then treated with 5nM R1881 or DMSO as a vehicle control (0nM R1881). VEGF mRNA expression was measured by qRT-PCR and normalized by β-actin levels as described in text. (B) VEGF mRNA expression in LNCaP cells was measured by qRT-PCR and normalized by 18S levels, as described in text. Cells were serum starved as described in A. For inhibition of androgen activity, cells were pre-treated with 10μM casodex for 2 hrs and then induced with 5nM R1881 for 24 hrs. Values represent fold change relative to DMSO treatment. A Student’s t-test was performed and significance was determined * (p < 0.05), ** (p < 0.01). (C) VEGF protein expression in LNCaP cells treated with 1nM R1881 for 0–48 hours. Protein expression was measured by western blotting as described in text, and β-actin levels were used as loading controls. (D) Cytoplasmic VEGF protein expression was measured by western blot of LNCaP cells treated as per (B). Image J analysis was performed and VEGF levels were normalized to β-actin levels. Shown are relative fold-changes in VEGF protein levels, normalized to β-actin and relative to untreated cells.Back to article page